λ Vectors for Stable Cloned Gene Expression

• Published in: Biotechnology progress Vol. 6; no. 4; pp. 277 - 282
• Main Authors: Padukone, N., Peretti, S. W., Ollis, D. F.
• Format: Journal Article
• Language: English
• Published: USA American Chemical Society 01.07.1990; American Institute of Chemical Engineers
• Subjects: Bacteriophage lambda - genetics; Bacteriophage lambda - physiology; beta-Galactosidase - biosynthesis; beta-Galactosidase - genetics; Biological and medical sciences; Biotechnology; Cloning, Molecular; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression; Genetic engineering; Genetic technics; Genetic Vectors; Lysogeny; Methods. Procedures. Technologies; Plasmids; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Stability; Escherichia coli; Inheritance; Lysogeny; β-D-Galactosidase; Gene expression; Virus; Plasmid; Lambda phage group; Enzymatic activity; Styloviridae; Production; Bacteria; Mutation; Phage lambda; Vector; Enterobacteriaceae; Phage
• ISSN: 8756-7938; 1520-6033
• DOI: 10.1021/bp00004a008

The bacteriophage λ offers a unique opportunity concurrently to minimize segregational instability in recombinant systems by chromosomal integration of the cloned gene and to achieve high cloned gene expression during an abortive lytic phase. Lysis leads approximately to a 100‐fold amplification of the cloned gene. Cell lysis in the lytic state is blocked by a specific mutation (Sam), allowing the cell to maintain its integrity, and λ DNA packaging is blocked by other mutations (Wam, Eam) that keep cloned genes open to transcription. In the presence of these mutations, extremely high levels of cloned β‐galactosidase (more than 15% of total cell protein) have been obtained during abortive lysis from vectors found to be essentially 100% stable for over 75 generations in the lysogenic phase.